云南师宗和江苏盱眙中华按蚊的分子鉴定及COII基因序列分析

Molecular Identification and COII gene analysis of Anopheles sinensis from Shizong County,Yunnan Province and Xuyi County,Jiangsu Province

为了了解云南省师宗县和江苏省盱眙县中华按蚊的分子鉴定及COII基因序列特征,2014年7月在云南省师宗县和江苏省盱眙县采集蚊虫标本,通过形态学鉴定出中华按蚊,再提取蚊虫基因组DNA,用COII基因特异引物和序列测定,然后采用生物信息学软件进行核苷酸序列同源性和遗传进化分析。结果在云南省师宗县县城周围采集蚊虫标本314只,其中按蚊234只(74.52%);在江苏省盱眙县桂五镇采集蚊虫标本212只,其中按蚊143只(67.45%)。经形态学鉴定,每个地方挑选了8只疑似中华按蚊进行序列分析,结果显示,16只蚊虫COII核苷酸序列同源性在98%-100%之间,遗传进化分析显示,16只蚊虫全部与中华按蚊位于同一进化分支内。表明了采用分子鉴定的方法可有效地进行蚊虫种类鉴定。

To understand the molecular taxonomy of An.sinensis and the characteristics of COII gene in Shizong County,Yunnan Province and Xuyi County,Jiangsu Province.In July 2014,we collected insects from Shizong County,Yunnan Province and Xuyi County,Jiangsu Province.We first identified An.sinensis by insect morphology.And then the genome DNA of mosquitos were extracted and amplified by COII gene specific primers and sequenced,which were analyzed by bioinformatics software for nucleotide sequence homology and phylogenetic relationships.We collected 314 insects from Shizong County,Yunnan Province,including 234 Anopheles（74.52 %,and 212 insects from Xuyi County,Jiangsu Province,including 143 Anopheles（67.45 %）.We identified them by morphology,and then selected 8 suspected An.sinensis from each of the two places for sequencing analysis.Results showed that the nucleotide sequence homology of all 16 insects COII genes varied from 98%-100%,and the phylogenetic analysis showed that all 16 insects and An.sinensis located in the same cluster.Molecular identification method is a very effective way to identify mosquito species.